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Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
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JoVE Journal Biologie
Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death

Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death

DOI:
10.3791/4442-v

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13:54 min

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October 21, 2012

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, ,
1Department of Molecular Microbiology, Faculty of Medicine,Technion – Israel Institute of Technology

Kapitel

  • 00:05Titel
  • 01:50Generation of Inducible Cell Lines
  • 06:19Induction of Knockdown and Transfection
  • 08:08DAPI Assay in Transfected Cells
  • 10:59Results: Detection of E4orf4-induced Cell Death in Inducible Cell Lines
  • 13:06Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

Contribution of the ACF chromatin remodeling factor to E4orf4-induced cell death was measured. The protocol includes selection of cell clones in which doxycycline treatment induces conditional knockdown of the ACF subunits Acf1 and SNF2h, and use of the DAPI assay to measure E4orf4-induced cell death in the inducible cell lines.

Tags

Conditional KnockdownGene ExpressionCell LinesACF Chromatin Remodeling FactorDoxycycline-inducible PromoterNeomycin Resistance GeneShRNA ConstructsAcf1SNF2hE4orf4 ProteinCell Death

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